GLUT1和PKM2在二甲双胍抑制人乳头状甲状腺癌细胞增殖中的作用  

GLUT1 and PKM2 involved in antitumoral effects of metformin in papillary thyroid cancer cells

作　　者：郭慧玲[1] 刘睿[1] 王红[1] 关海霞[1] 单忠艳[1] 滕卫平[1] 

机构地区：[1]中国医科大学附属第一医院内分泌科,内分泌研究所,辽宁省重点实验室,辽宁沈阳110001

出　　处：《现代肿瘤医学》2015年第7期900-904,共5页Journal of Modern Oncology

基　　金：中华医学会临床医学科研专项资金-施贵宝内分泌糖尿病研究项目(编号:13040480433);辽宁省高等学校创新团队项目(编号:LT2012015);沈阳市科学技术项目计划(编号:F11-244-1-00)

摘　　要：目的:观察不同浓度二甲双胍对人乳头状甲状腺癌(papillary thyroid cancer,PTC)细胞增殖、凋亡的影响,检测细胞中葡萄糖转运关键蛋白—葡萄糖转运体1(GLUT1)和糖酵解关键酶—M2型丙酮酸激酶(PKM2)表达水平的变化。方法:体外培养人PTC细胞BCPAP和K1,分为对照组和不同浓度二甲双胍(1、5和10mmol/L)处理组,并分别培养24h、48h和72h。四甲基偶氮唑蓝(MTT)法检测细胞增殖,Annexin VFITC/PI流式细胞术检测细胞凋亡,实时定量PCR检测GLUT1和PKM2 mRNA的表达,Western blot检测GLUT1和PKM2蛋白的表达。结果:与对照组相比,二甲双胍抑制PTC细胞增殖,随浓度及处理时间的增加,抑制效应加大,呈剂量-时间依赖性(P<0.05)。二甲双胍诱导PTC细胞凋亡,随浓度及处理时间的增加,其中晚期凋亡率明显增加,呈剂量-时间依赖性(P<0.05)。二甲双胍可抑制PTC细胞中GLUT1和PKM2mRNA的表达水平,呈剂量-时间依赖性(P<0.05)。二甲双胍可抑制PTC细胞中GLUT1和PKM2蛋白的表达水平,呈时间依赖性。结论:二甲双胍抑制人PTC细胞增殖并诱导凋亡,其机制之一可能是抑制葡萄糖转运关键蛋白和糖酵解关键酶的表达,负面影响癌细胞生长的能量供应。Objective： To investigate the effects of different concentration of metformin on the proliferation and apoptosis of papillary thyroid cancer（ PTC）,and to determine whether metformin causes alterations of both GLUT1 and PKM2 levels. Methods： After treating with metformin（ 0,1,5,10 mmol / L）,the viability of PTC cell lines（ BCPAP,K1） was analyzed by 3-（ 4,5- dimethyl- thiazol- 2- yl）- 2,5- diphenyltetrazolium bromide（ MTT）. Cell cycle analysis was performed by Annexin V- FITC / PI flow cytometry,real- time polymerase chain reaction and Western blot were performed to determine the expression of GLUT1 and PKM2. Results： Compared with control group,MTT assays showed that the proliferation of PTC cell lines was inhibited in a dose- time dependent manner after treating with metformin（ P〈0. 05）. Moreover,metformin induced the late apoptosis of the PTC cell lines（ P〈0. 05）. The relative mRNA levels of GLUT1 and PKM2 were inhibited in a dose- time dependent manner by metformin（ P〈0. 05）. The protein expression of GLUT1 and PKM2 in PTC cells was decreased by metformin in a time dependent manner. Conclusion： Metformin provides an antitumoral effect in PTC cells. This effect may be mediated through a decrease of GLUT1 and PKM2 expression which leading to an energy crisis in cancer cells.

关 键 词：二甲双胍 人乳头状甲状腺癌细胞 GLUT1 PKM2 

分 类 号：R736.1[医药卫生—肿瘤]